Engineering ligand-specific biosensors for aromatic amino acids and neurochemicals.
Engineering ligand-specific biosensors for aromatic amino acids and neurochemicals.
复制标题
DOI：
10.1016/j.cels.2021.10.006
复制
发表时间：
2022-03-16
期刊：
影响因子：
9.3
通讯作者：
Moon TS
中科院分区：
文献类型：
--
作者：
Rottinghaus AG;Xi C;Amrofell MB;Yi H;Moon TS
微生物生物传感器在代谢工程和医学中有着广泛的应用。化学浓度的特异性和准确定量允许酶途径的适应性调节和诊断报告基因的时间精确表达。虽然生物传感器应该区分具有不同生物功能的结构相似的配体，但这种特定的传感器在自然界中很少发现，并且具有挑战性。使用大肠coli Nissle 1917，一种被普遍认为是安全的微生物，我们表征了两种混杂识别芳香族氨基酸或神经化学物质的生物传感器系统。为了提高传感器的选择性和灵敏度，我们应用合理的蛋白质工程，通过识别和诱变氨基酸残基，并成功地展示了苯丙氨酸，酪氨酸，苯乙胺和酪胺的配体特异性生物传感器。此外，我们的方法揭示了FeaR调节剂的未表征结构，包括配体结合中的关键残基。这些结果为开发用于不同应用的动力学适应性微生物奠定了基础。这项工作代表了蛋白质工程的一项重大成就，包括芳香族氨基酸（如苯丙氨酸和酪氨酸）以及神经化学物质（如苯乙胺和酪胺）的配体特异性传感器，它们在结构上相似，并与各种医疗条件有关。它为开发具有高度特异性的医学相关益生菌和可穿戴传感器奠定了基础，这对于区分具有不同功能的代谢物以及创建用于准确诊断的智能益生菌和无细胞生物传感器至关重要。
Microbial biosensors have diverse applications in metabolic engineering and medicine. Specific and accurate quantification of chemical concentrations allows for adaptive regulation of enzymatic pathways and temporally-precise expression of diagnostic reporters. Although biosensors should differentiate structurally similar ligands with distinct biological functions, such specific sensors are rarely found in nature and challenging to create. Using E. coli Nissle 1917, a generally regarded as safe microbe, we characterized two biosensor systems that promiscuously recognize aromatic amino acids or neurochemicals. To improve the sensors’ selectivity and sensitivity, we applied rational protein engineering by identifying and mutagenizing amino acid residues and successfully demonstrated the ligand-specific biosensors for phenylalanine, tyrosine, phenylethylamine, and tyramine. Additionally, our approach revealed insights into the uncharacterized structure of the FeaR regulator, including critical residues in ligand binding. These results lay the groundwork for developing kinetically-adaptive microbes for diverse applications. This work represents a considerable achievement in protein engineering and includes the ligand-specific sensors for aromatic amino acids such as phenylalanine and tyrosine as well as neurochemicals such as phenylethylamine and tyramine, which are structurally similar and have been implicated in a variety of medical conditions. It lays the groundwork for developing medically-relevant probiotic and wearable sensors with high specificity, which is critical to differentiate metabolites with divergent functions and create smart probiotics and cell-free biosensors for accurate diagnostics.
登录
查看更多内容
影响因子：
5
作者：
Mahr, Regina;von Boeselager, Raphael Freiherr;Frunzke, Julia
通讯作者：
Frunzke, Julia
影响因子：
14.8
作者：
Meyer, Adam J.;Segall-Shapiro, Thomas H.;Voigt, Christopher A.
通讯作者：
Voigt, Christopher A.
影响因子：
14.9
作者：
Lee YJ;Hoynes-O'Connor A;Leong MC;Moon TS
通讯作者：
Moon TS
影响因子：
14.8
作者：
Combs SA;Deluca SL;Deluca SH;Lemmon GH;Nannemann DP;Nguyen ED;Willis JR;Sheehan JH;Meiler J
通讯作者：
Meiler J
影响因子：
158.5
作者：
MITCHELL, G;LAROCHELLE, J;DALLAIRE, L
通讯作者：
DALLAIRE, L